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Justo Lukban y Rilles was born on May 28, 1863, the second child of a wealthy couple, Don Agustin Lukban and Dona Andrea Rilles, in Labo, Ambos Camarines, now the province of Camarines Norte. He had three brothers, Cayetano, Miguel, and Vicente, and two sisters, Rafaela and Concha.

After the completion of his elementary education, he enrolled in 1873 at the Colegio de San Juan de Letran for his Bachelor of Arts. Then he took up medicine at the University of Santo Tomas. He held the position of Ayundante Director of the School of Medicine in 1884. In 1888, he was conferred the Licentiate of Medicine, subsequently establishing a clinic in Manila.